Kateřina Abrhámová, Martina Groušlová, Anna Valentová, Xinxin Hao, Beidong Liu, Martin Převorovský, Ondřej Gahura, František Půta, Per Sunnerhagen, Petr Folk
{"title":"截断剪接体 \"绳索蛋白 \"Prp45会导致依赖于Htz1的表型。","authors":"Kateřina Abrhámová, Martina Groušlová, Anna Valentová, Xinxin Hao, Beidong Liu, Martin Převorovský, Ondřej Gahura, František Půta, Per Sunnerhagen, Petr Folk","doi":"10.1080/15476286.2024.2348896","DOIUrl":null,"url":null,"abstract":"<p><p>Spliceosome assembly contributes an important but incompletely understood aspect of splicing regulation. Prp45 is a yeast splicing factor which runs as an extended fold through the spliceosome, and which may be important for bringing its components together. We performed a whole genome analysis of the genetic interaction network of the truncated allele of <i>PRP45</i> (<i>prp45</i>(1-169)) using synthetic genetic array technology and found chromatin remodellers and modifiers as an enriched category. In agreement with related studies, H2A.Z-encoding <i>HTZ1</i>, and the components of SWR1, INO80, and SAGA complexes represented prominent interactors, with <i>htz1</i> conferring the strongest growth defect. Because the truncation of Prp45 disproportionately affected low copy number transcripts of intron-containing genes, we prepared strains carrying intronless versions of <i>SRB2</i>, <i>VPS75</i>, or <i>HRB1</i>, the most affected cases with transcription-related function. Intron removal from <i>SRB2</i>, but not from the other genes, partly repaired some but not all the growth phenotypes identified in the genetic screen. The interaction of <i>prp45</i>(1-169) and <i>htz1</i>Δ was detectable even in cells with <i>SRB2</i> intron deleted (<i>srb2</i>Δi). The less truncated variant, <i>prp45</i>(1-330), had a synthetic growth defect with <i>htz1</i>Δ at 16°C, which also persisted in the <i>srb2</i>Δi background. Moreover, <i>htz1</i>Δ enhanced <i>prp45</i>(1-330) dependent pre-mRNA hyper-accumulation of both high and low efficiency splicers, genes <i>ECM33</i> and <i>COF1</i>, respectively. We conclude that while the expression defects of low expression intron-containing genes contribute to the genetic interactome of <i>prp45</i>(1-169), the genetic interactions between <i>prp45</i> and <i>htz1</i> alleles demonstrate the sensitivity of spliceosome assembly, delayed in <i>prp45</i>(1-169), to the chromatin environment.</p>","PeriodicalId":21351,"journal":{"name":"RNA Biology","volume":null,"pages":null},"PeriodicalIF":3.6000,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11085953/pdf/","citationCount":"0","resultStr":"{\"title\":\"Truncating the spliceosomal 'rope protein' Prp45 results in Htz1 dependent phenotypes.\",\"authors\":\"Kateřina Abrhámová, Martina Groušlová, Anna Valentová, Xinxin Hao, Beidong Liu, Martin Převorovský, Ondřej Gahura, František Půta, Per Sunnerhagen, Petr Folk\",\"doi\":\"10.1080/15476286.2024.2348896\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Spliceosome assembly contributes an important but incompletely understood aspect of splicing regulation. Prp45 is a yeast splicing factor which runs as an extended fold through the spliceosome, and which may be important for bringing its components together. We performed a whole genome analysis of the genetic interaction network of the truncated allele of <i>PRP45</i> (<i>prp45</i>(1-169)) using synthetic genetic array technology and found chromatin remodellers and modifiers as an enriched category. In agreement with related studies, H2A.Z-encoding <i>HTZ1</i>, and the components of SWR1, INO80, and SAGA complexes represented prominent interactors, with <i>htz1</i> conferring the strongest growth defect. Because the truncation of Prp45 disproportionately affected low copy number transcripts of intron-containing genes, we prepared strains carrying intronless versions of <i>SRB2</i>, <i>VPS75</i>, or <i>HRB1</i>, the most affected cases with transcription-related function. Intron removal from <i>SRB2</i>, but not from the other genes, partly repaired some but not all the growth phenotypes identified in the genetic screen. The interaction of <i>prp45</i>(1-169) and <i>htz1</i>Δ was detectable even in cells with <i>SRB2</i> intron deleted (<i>srb2</i>Δi). The less truncated variant, <i>prp45</i>(1-330), had a synthetic growth defect with <i>htz1</i>Δ at 16°C, which also persisted in the <i>srb2</i>Δi background. Moreover, <i>htz1</i>Δ enhanced <i>prp45</i>(1-330) dependent pre-mRNA hyper-accumulation of both high and low efficiency splicers, genes <i>ECM33</i> and <i>COF1</i>, respectively. We conclude that while the expression defects of low expression intron-containing genes contribute to the genetic interactome of <i>prp45</i>(1-169), the genetic interactions between <i>prp45</i> and <i>htz1</i> alleles demonstrate the sensitivity of spliceosome assembly, delayed in <i>prp45</i>(1-169), to the chromatin environment.</p>\",\"PeriodicalId\":21351,\"journal\":{\"name\":\"RNA Biology\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":3.6000,\"publicationDate\":\"2024-01-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11085953/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"RNA Biology\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://doi.org/10.1080/15476286.2024.2348896\",\"RegionNum\":3,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2024/5/6 0:00:00\",\"PubModel\":\"Epub\",\"JCR\":\"Q2\",\"JCRName\":\"BIOCHEMISTRY & MOLECULAR BIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"RNA Biology","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1080/15476286.2024.2348896","RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2024/5/6 0:00:00","PubModel":"Epub","JCR":"Q2","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
Truncating the spliceosomal 'rope protein' Prp45 results in Htz1 dependent phenotypes.
Spliceosome assembly contributes an important but incompletely understood aspect of splicing regulation. Prp45 is a yeast splicing factor which runs as an extended fold through the spliceosome, and which may be important for bringing its components together. We performed a whole genome analysis of the genetic interaction network of the truncated allele of PRP45 (prp45(1-169)) using synthetic genetic array technology and found chromatin remodellers and modifiers as an enriched category. In agreement with related studies, H2A.Z-encoding HTZ1, and the components of SWR1, INO80, and SAGA complexes represented prominent interactors, with htz1 conferring the strongest growth defect. Because the truncation of Prp45 disproportionately affected low copy number transcripts of intron-containing genes, we prepared strains carrying intronless versions of SRB2, VPS75, or HRB1, the most affected cases with transcription-related function. Intron removal from SRB2, but not from the other genes, partly repaired some but not all the growth phenotypes identified in the genetic screen. The interaction of prp45(1-169) and htz1Δ was detectable even in cells with SRB2 intron deleted (srb2Δi). The less truncated variant, prp45(1-330), had a synthetic growth defect with htz1Δ at 16°C, which also persisted in the srb2Δi background. Moreover, htz1Δ enhanced prp45(1-330) dependent pre-mRNA hyper-accumulation of both high and low efficiency splicers, genes ECM33 and COF1, respectively. We conclude that while the expression defects of low expression intron-containing genes contribute to the genetic interactome of prp45(1-169), the genetic interactions between prp45 and htz1 alleles demonstrate the sensitivity of spliceosome assembly, delayed in prp45(1-169), to the chromatin environment.
期刊介绍:
RNA has played a central role in all cellular processes since the beginning of life: decoding the genome, regulating gene expression, mediating molecular interactions, catalyzing chemical reactions. RNA Biology, as a leading journal in the field, provides a platform for presenting and discussing cutting-edge RNA research.
RNA Biology brings together a multidisciplinary community of scientists working in the areas of:
Transcription and splicing
Post-transcriptional regulation of gene expression
Non-coding RNAs
RNA localization
Translation and catalysis by RNA
Structural biology
Bioinformatics
RNA in disease and therapy